CN106834419A - A kind of method that use excised leaf identifies sugarcane toppers rot-resistant - Google Patents
A kind of method that use excised leaf identifies sugarcane toppers rot-resistant Download PDFInfo
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- CN106834419A CN106834419A CN201710132453.3A CN201710132453A CN106834419A CN 106834419 A CN106834419 A CN 106834419A CN 201710132453 A CN201710132453 A CN 201710132453A CN 106834419 A CN106834419 A CN 106834419A
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Abstract
The invention discloses a kind of method that use excised leaf identifies sugarcane toppers rot-resistant, comprise the following steps：(1) sugarcane top rot bacterium is seeded on PDA culture medium flat board, 25 28 DEG C are cultivated 35 days；(2) fresh cane blade is gathered, blade is cut into fragment by cleaning blade, and vein both sides are stabbed in the middle of blade, and the bacterium block for taking sugarcane top rot bacterium is seeded in puncture wounds, wound is attached to the one side that length has mycelia；(3) Sugarcane Leaves that will connect bacterium are placed in culture dish, spray sterilized water, in placement 35 days under room temperature condition；(4) length of Disease investigation, observation and measurement scab, along the length of vein orientation measurement yellow halo, evaluates the top rot resistance of sugar cane breed according to scab length.Authentication method of the invention, the plantation without carrying out cane seedling according to season, is not limited by sugarcane seedling age and place, easy to operate, reproducible, for the screening of great amount of samples have preferable saving of labor, it is time saving the characteristics of.
Description
Technical field
The present invention relates to a kind of method that use excised leaf identifies sugarcane toppers rot-resistant.
Background technology
Sugarcane top rot is a kind of important leaf diseases in sugarcane production, sugarcane top rot (Pokkah Boeng) be by
A kind of various microbial sugarcane leaf diseases of reaping hook, pathogen is gibberella fujikuroi germ aggregate species (Gibberella
Fujikuroi complex), Invisible element at least include Fuarium sacchri, F.proliferatum,
5 kinds of sickle-like bacteria such as F.verticillioides, F.subglutinan and F.andiyazi.Pathogen is mainly propagated by air-flow
Infected to sugarcane lobus cardiacus and caused harm, cause blade dead, plant strain growth is obstructed, can seriously cause plant dead, cause the sugarcane underproduction
5%-20%, cane sugar reduction causes to treat that influence is more significantly up to 3% on susceptible sugar cane breed.The disease is nearly all sweet
Sugarcane production countries and regions generally occur, although carrying out prevention and control by chemical agent can reduce the generation of the disease.But application is anti-
Sick kind is effectively reduced the generation of sugarcane top rot, and reduces pollution of the chemical pesticide to environment.
In recent years, sugarcane top rot in the generation of China's sugarcane district in the trend that gradually aggravates, sugarcane production to China and
Sugar industry sustainable development constitutes a serious threat.Therefore, a set of fast and accurate sugarcane toppers rot-resistant authentication method is set up, is had
Scientific evaluation is carried out beneficial to existing sugarcane germplasm and kind, is the breeding for disease resistance of sugarcane top rot and the sum of kind
Rational deployment provides scientific basis.
Evaluation of resistance of the artificial stab inoculation of Seedling of Sugarcane Under Water Stress to top rot is had in the prior art, but whole process needs
The sterilization of experience sugarcane kind, sowing, could be inoculated with after after long to 3~4 complete leaves of sugarcane seedling, need to see after inoculation within 10~15 days
Examine and judge the disease resistance of sugar cane breed.Whole cycle is more long, and sugarcane is at least needed from being seeded into the appropriate seedling age that can be inoculated with
The time of 1.5~2 months, and greenhouse humiture whard to control, different season sowings, the resistance of sugarcane are different, easily cause
Replica test result it is inconsistent.In addition, when being inoculated with not in time, with the increase of seedling age, kind resistance meeting in itself
Improve constantly, inoculation is difficult to cause Sugarcane Leaves to be fallen ill, and causes the differentiation error of resistance level.
The information for being disclosed in the background section is merely intended to increase the understanding to general background of the invention, without answering
In being considered as recognizing or imply in any form that the information structure has been the prior art well known to persons skilled in the art.
The content of the invention
It is an object of the invention to provide a kind of method that use excised leaf identifies sugarcane toppers rot-resistant, the method energy
Shorten the cycle of characterization and evaluation, simple, accurate, effective, reproducible, the collection of sample is difficult to receive cultivation season and seedling age
Influence.
To achieve the above object, the invention provides a kind of method that use excised leaf identifies sugarcane toppers rot-resistant,
Comprise the following steps：
(1) sugarcane top rot bacterium is seeded on PDA culture medium flat board, 25-28 DEG C is cultivated 3-5 days；
(2) fresh cane blade is gathered, blade is cut into fragment by cleaning blade, and vein both sides are stabbed in the middle of blade, are taken
The bacterium block of sugarcane top rot bacterium is seeded in puncture wounds, and wound is attached to the one side that length has mycelia；
(3) Sugarcane Leaves that will connect bacterium are placed in culture dish, spray sterilized water, in placement 3-5 days under room temperature condition；
(4) length of Disease investigation, observation and measurement scab, along the length of vein orientation measurement yellow halo, according to
Scab length is evaluated sugar cane breed top rot resistance, a peak and a minimum is removed, with remaining numerical value
Average value represents the resistance of the sugar cane breed；The morbidity grading standard of sugarcane top rot is as follows：
0 grade：Without disease symptom；
1 grade：Acupuncture wound dark brown, yellow halo is along vein direction length≤1.0cm；
2 grades：Acupuncture wound dark brown, yellow halo is along vein direction length 1.1-2.0cm；
3 grades：Acupuncture wound dark brown, yellow halo is along vein direction length 2.1-3.0cm；
4 grades：Acupuncture wound dark brown, yellow halo is along vein direction length 3.1-4.0cm；
5 grades：Acupuncture wound dark brown, yellow halo is along vein direction length>4.0cm.
Preferably, in above-mentioned technical proposal, the morbidity grading standard according to sugarcane top rot is to sugarcane resistance class
Divided, the sugarcane resistance class criteria for classifying is as follows：
Height is anti-：Morbidity grade is 0；
It is disease-resistant：Morbidity grade is 1；
In resist：Morbidity grade is 2；
Middle sense：Morbidity grade is 3；
It is susceptible：Morbidity grade is 4
Height sense：Morbidity grade is 5.
Preferably, in above-mentioned technical proposal, collection fresh cane blade is in the step (2), from lobus cardiacus down number the 2nd
Blade is fully deployed to the 4th.
Preferably, in above-mentioned technical proposal, the fragment length that step (2) Leaf is cut into is 10-14cm.
Preferably, taking bacterium block in above-mentioned technical proposal, in the step (2) is, bacterium is taken with the pipettor gun head of sterilizing
Block, a diameter of 5-6mm of bacterium block.
Preferably, in above-mentioned technical proposal, the amount of sprinkling sterilized water is 3-5ml in the step (3).
Preferably, in above-mentioned technical proposal, culture dish selects a diameter of 15cm in the step (3).
Compared with prior art, the present invention has the advantages that：It is of the invention a kind of using excised leaf identification sugarcane
The method of top rot resistance, sugar cane breed is determined to top rot resistance level, the collection of Sugarcane Leaves sample using Vitro Inoculation Technique
Sugarcane seedling age is not received, the influence in period is planted, condition determination is easy to control.Sugarcane excised leaf can be one under the conditions of heat and moisture preserving
Do not dry up in fixing time, it is ensured that pathogen is infected.The preparation saving of labor of inoculum is saved trouble, and is directly seeded in PDA culture medium and is put down
Plate incubated at room temperature, liquid medium and conidium liquid are made without extra.Without carrying out cane seedling according to season
Plantation, also do not limited by sugarcane seedling age and place, it is easy to operate indoors, it is reproducible, particularly with great amount of samples
Screening have preferable saving of labor, it is time saving the characteristics of.
Specific embodiment
With reference to specific embodiment, specific embodiment of the invention is described in detail, it is to be understood that this hair
Bright protection domain is not limited by specific embodiment.
Embodiment
A kind of method that use excised leaf identifies sugarcane toppers rot-resistant, comprises the following steps：
1st, the preparation of inoculum
Cultivated respectively using two stronger sugarcane top rot sickle-like bacteria bacterial strains of pathogenicity.Specially：Respectively will be sweet
Sugarcane top rot pathogen is seeded on PDA culture medium flat board and activates 3 days, the new PDA culture medium flat board of picking edge mycelium inoculation
On, 25-28 DEG C is cultivated 3-5 days, and mycelia block (diameter about 5-6mm) is taken with the 1mL pipettor gun heads (one end of wide opening) of sterilizing,
It is stand-by.
2nd, the treatment of inoculation material
Show as sugarcane top rot susceptible variety GT37 and LC05-136 in selection production, disease-resistant variety ROC22 and 5
Test product system of area (G505-3217, GR2, GT08-6, GT09-1743 and GT08-120), collection fresh cane blade is (past from lobus cardiacus
Lower number the 2nd to the 4th is fully deployed blade), rinse the dust on surface well with running water, dry, it is cut into the fragment of about 12cm.
3rd, it is inoculated with
With pin, vein both sides are stabbed in the middle of blade, and 4 pins are pierced altogether according to the mode of about 3 × 3mm.Take sugarcane top rot
Bacterium bacterium block, is seeded in puncture wounds, and wound is attached to the one side that length has mycelia, and bacterium dish is completely covered stabs mouth.Each bacterial strain
Respectively in 10 side inoculations of blade vein, opposite side is to connect pure PDA culture medium as control.To connect the Sugarcane Leaves of bacterium as
In the glass culture dish of diameter 15cm, appropriate sterilized water (about 3-5mL) is sprayed.In placing 3-5 under room temperature condition (25-28 DEG C)
My god.
4th, Disease investigation
Observation and the length of measurement scab, along the length of vein orientation measurement yellow halo, according to scab length to sweet
Sugarcane kind top rot resistance is evaluated, and removes a peak and a minimum, and being represented with the average value of remaining numerical value should
The resistance of sugar cane breed；To stab inoculation, the morbidity grading standard of sugarcane top rot is as follows：
0 grade：Without disease symptom；
1 grade：Acupuncture wound dark brown, yellow halo is along vein direction length≤1.0cm；
2 grades：Acupuncture wound dark brown, yellow halo is along vein direction length 1.1-2.0cm；
3 grades：Acupuncture wound dark brown, yellow halo is along vein direction length 2.1-3.0cm；
4 grades：Acupuncture wound dark brown, yellow halo is along vein direction length 3.1-4.0cm；
5 grades：Acupuncture wound dark brown, yellow halo is along vein direction length>4.0cm.
Morbidity grading standard according to sugarcane top rot is divided to sugarcane resistance class, and sugarcane resistance class is drawn
Minute mark is accurate as follows：
Height is anti-：Morbidity grade is 0；
It is disease-resistant：Morbidity grade is 1；
In resist：Morbidity grade is 2；
Middle sense：Morbidity grade is 3；
It is susceptible：Morbidity grade is 4
Height sense：Morbidity grade is 5.
5th, resistance result identification
Different sugar cane breed inoculation top rot bacterium resistance level qualification results are shown in Table 1.Different sugar cane breeds are to the different tips
The susceptible situation of maize ear rot bacterial strain is different, and it is the stronger bacterial strain of pathogenicity respectively to 8 sugar cane breeds to use the screening of two early stages
(material) is inoculated with, and except some differences of performance resistance on respective material, but aggregate level is more consistent.For example, resistance
When two worst sugar cane breeds LC05-136 and GT37 are inoculated with 2 pathogenic strains respectively, resistance is shown as worst；And resistance
When stronger Sugarcane smut GT08-120 is inoculated with 2 pathogenic strains respectively, preferable resistance is also shown as.Indoor in vitro resistance
Qualification result is basically identical with the performance for producing.Illustrate that the method can preferably suitable for the sieve of sugarcane toppers rot-resistant material
Choosing.
The different sugar cane breed inoculation top rot bacterium resistance level qualification results of table 1
The foregoing description to specific illustrative embodiment of the invention be in order to illustrate and illustration purpose.These descriptions
It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to above-mentioned teaching, can be much changed
And change.The purpose of selecting and describing the exemplary embodiment is that explaining that certain principles of the invention and its reality should
With so that those skilled in the art can realize and using a variety of exemplaries of the invention and
A variety of selections and change.The scope of the present invention is intended to be limited by claims and its equivalents.
Claims (7)
1. a kind of method that use excised leaf identifies sugarcane toppers rot-resistant, it is characterised in that comprise the following steps：
(1) sugarcane top rot bacterium is seeded on PDA culture medium flat board, 25-28 DEG C is cultivated 3-5 days；
(2) fresh cane blade is gathered, blade is cut into fragment by cleaning blade, and vein both sides are stabbed in the middle of blade, take sugarcane
The bacterium block of top rot bacterium is seeded in puncture wounds, and wound is attached to the one side that length has mycelia；
(3) Sugarcane Leaves that will connect bacterium are placed in culture dish, spray sterilized water, in placement 3-5 days under room temperature condition；
(4) length of Disease investigation, observation and measurement scab, along the length of vein orientation measurement yellow halo, according to scab
Length is evaluated sugar cane breed top rot resistance, a peak and a minimum is removed, with the average of remaining numerical value
Value represents the resistance of the sugar cane breed；The morbidity grading standard of sugarcane top rot is as follows：
0 grade：Without disease symptom；
1 grade：Acupuncture wound dark brown, yellow halo is along vein direction length≤1.0cm；
2 grades：Acupuncture wound dark brown, yellow halo is along vein direction length 1.1-2.0cm；
3 grades：Acupuncture wound dark brown, yellow halo is along vein direction length 2.1-3.0cm；
4 grades：Acupuncture wound dark brown, yellow halo is along vein direction length 3.1-4.0cm；
5 grades：Acupuncture wound dark brown, yellow halo is along vein direction length>4.0cm.
2. the method that use excised leaf according to claim 1 identifies sugarcane toppers rot-resistant, it is characterised in that according to
The morbidity grading standard of sugarcane top rot is divided to sugarcane resistance class, and the sugarcane resistance class criteria for classifying is as follows：
Height is anti-：Morbidity grade is 0；
It is disease-resistant：Morbidity grade is 1；
In resist：Morbidity grade is 2；
Middle sense：Morbidity grade is 3；
It is susceptible：Morbidity grade is 4
Height sense：Morbidity grade is 5.
3. the method that use excised leaf according to claim 1 identifies sugarcane toppers rot-resistant, it is characterised in that described
Collection fresh cane blade is that down number the 2nd to the 4th is fully deployed blade from lobus cardiacus in step (2).
4. it is according to claim 1 determine sugarcane top rot germ pathogenicity method, it is characterised in that the step
(2) fragment length that Leaf is cut into is 10-14cm.
5. it is according to claim 1 determine sugarcane top rot germ pathogenicity method, it is characterised in that the step
(2) bacterium block is taken in is, bacterium block, a diameter of 5-6mm of bacterium block are taken with the pipettor gun head of sterilizing.
6. it is according to claim 1 determine sugarcane top rot germ pathogenicity method, it is characterised in that the step
(3) amount of sprinkling sterilized water is 3-5ml in.
7. it is according to claim 1 determine sugarcane top rot germ pathogenicity method, it is characterised in that the step
(3) culture dish selects a diameter of 15cm in.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710132453.3A CN106834419A (en) | 2017-03-07 | 2017-03-07 | A kind of method that use excised leaf identifies sugarcane toppers rot-resistant |
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| CN201710132453.3A CN106834419A (en) | 2017-03-07 | 2017-03-07 | A kind of method that use excised leaf identifies sugarcane toppers rot-resistant |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108486033A (en) * | 2018-02-23 | 2018-09-04 | 云南省德宏热带农业科学研究所 | A kind of in-vitro verification method of coffee germplasm rust resistance |
| CN109136327A (en) * | 2018-10-08 | 2019-01-04 | 华中农业大学 | A method of identification citrus plant Resistance To Root Rot Disease |
| CN112430639A (en) * | 2020-11-20 | 2021-03-02 | 广西大学 | Method for determining pathogenicity of diaphorina citri endophytic fungi |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104450862A (en) * | 2014-11-04 | 2015-03-25 | 广西大学 | Sugarcane top rot resistance identification method |
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2017
- 2017-03-07 CN CN201710132453.3A patent/CN106834419A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104450862A (en) * | 2014-11-04 | 2015-03-25 | 广西大学 | Sugarcane top rot resistance identification method |
Non-Patent Citations (3)
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| 张玉娟: "甘蔗梢腐病病原分子检测及甘蔗组合、品种的抗病性评价", 《中国优秀硕士学位论文全文数据库》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108486033A (en) * | 2018-02-23 | 2018-09-04 | 云南省德宏热带农业科学研究所 | A kind of in-vitro verification method of coffee germplasm rust resistance |
| CN109136327A (en) * | 2018-10-08 | 2019-01-04 | 华中农业大学 | A method of identification citrus plant Resistance To Root Rot Disease |
| CN112430639A (en) * | 2020-11-20 | 2021-03-02 | 广西大学 | Method for determining pathogenicity of diaphorina citri endophytic fungi |
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Application publication date: 20170613 |